Study On The Migration Parameters Of Target Proteins In Multi-Compartment Electrophoresis And Screening Of Active Components Of Scutellaria Baicalensis

Traditional Chinese medicine（TCM）is an important part of the world medicine field,the embodiment of the TCM play efficacy is multicomponent acting on multiple targets in multiple signaling pathways play a synergy,can effectively improve efficacy and reduce side effects,but the TCM composition is complicated and varied,give play to the role of pharmacological composition is not clear,synergy mechanism is not clear,so TCM active ingredients research is an essential part of modern Chinese medicine,in which the key for the screening of active ingredients.The existing screening techniques,such as Ultrafiltration,Balanced dialysis and Immobilized enzyme,have been proved to be feasible for the screening of active ingredients,but they all have some limitations.Therefore,Our laboratory has developed Multi-compartment electrophoresis technology in the early stage,and it is feasible to realize rapid screening of potential active ingredients from natural products by Multi-compartment electrophoresis（MCESS）combined with liquid-mass spectrometry（HPLC-MS/MS）technology.The technology is fast,accurate,and screening of mild conditions,to maintain the advantages of three dimensional structure of the target protein.The application of Multi-compartment electrophoresis needs to be improved.Since the selected active components were dissociated from the target protein/active component complex,the migration of target protein was directly related to the component from screening to activity,which affected the accuracy and reproducibility of screening active compounds.Therefore,to explore the correlation between parameters such as voltage,buffer solution type,ionic strength,p H and time and the amount of target protein migration,to determine the best experimental parameters and to develop reference experimental conditions for target protein,It was applied to screen potentialα-glucosidase inhibitors in Scutellaria baicalensis,and to provide a good foundation for the efficient application of Multi-compartment electrophoresis technology screening in the future.The research contents and conclusions are as follows:Target protein migration parameters in Mult-icompartment electrophoresis,including voltage,buffer solution type,ionic strength,p H,electrophoresis time,explores the human serum albumin,trypsin,the migration of tyrosinase and the correlation between the parameters of the final and the best conditions for experiments can be used as target protein migration parameter selection on the basis of future experiments,high efficiency for the future application of Mult-icompartment electrophoresis technology provide the basis for a screening of the active ingredient.Based on the previous studies,the experimental conditions ofα-glucosidase migration were as follows:applying voltage was 12 V,buffer solution was ammonium acetate aqueous solution,ionic strength was 0.01 M,buffer solution p H was 4.0,and electrophoresis time was 8 min.By calculation,the migration amount is 50.7%,which is nearly twice as much as the previous method,and the electrophoresis time is nearly twice as much.In this paper,α-glucosidase was taken as the research object,through MCESS combine HPLC-MS/MS,successful from the extract of Scutellaria baicalensis screening to seven potentialα-glucosidase inhibitor.In vitro inhibition experiments showed that Baicalin and Wogonoside had good inhibitory effect onα-glucosidase,which were similar to acarbose,with IC₅₀values of 62.02±4.134μmol^-1and80.04±4.628μmol^-1.The results of this study can provide a basis for the further study of the hypoglycemic mechanism of Scutellaria baicalensis and the screening ofα-glucosidase inhibitors from natural products in the future.